The Targeted Long-acting Combination Antiretroviral Therapy (TLC-ART) Program has discovered a novel platform that can stabilize insoluble and soluble antiretroviral drugs together in a nanosuspension which provides long-acting (LA) plasma and cell concentrations in pre-clinical studies. The lead formulation (called TLC-ART 101) combines three FDA-approved drugs (active pharmaceutical ingredients, APIs) that are available generically lopinavir (LPV), ritonavir (RTV) and tenofovir (TFV). While TLC-ART 101 may not be developed as a commercial product, substantial pre-clinical data exists and the program has received favorable regulatory pathway guidance from the FDA. TLC-ART 101 is years ahead of other TLC-ART formulations in development. In this application, we propose a first-in-human, hypothesis-driven, clinical trial to provide human data about this novel, next generation, LA drug-combination platform technology. The trial will address multiple questions about the human pharmacokinetics (PK) and safety of this new platform. The project will be managed by an experienced, multi-disciplinary Study Leadership Team. The clinical trial will conducted by the staff at the UW AIDS Clinical Trials Unit (a Division of AIDS-approved research site affiliated with the AIDS Clinical Trials Group), overseen by an independent Data Monitoring Committee, and monitored by independent monitors. The proposed trial is an open-label, single-arm study with a pharmacologically- guided dose and duration adaptive design. The trial's hypothesis is that the TLC-ART drug-combination nanoparticle platform is safe and able to transform the PK of three short-acting generic and physically diverse antiretroviral drugs to create a LA concentration-time course in human plasma and cells. The primary objectives of the clinical trial are: 1) To characterize the plasma concentration-time course and PK of a single dose of the three APIs of TLC-ART 101 (LPV, RTV, and TFV) administered by sub-cutaneous injection, and 2) To characterize the safety and tolerability of a single subcutaneous injection of TLC-ART 101. The trial also has 4 exploratory mechanistic objectives that will provide information about the cell-targeting characteristics of TLC-ART 101 (meaning intracellular blood and lymphoid cells have equal or higher API concentrations than plasma), as well as exploring whether there are sex differences in the PK parameters of the 3 APIs. Twelve to 16 healthy persons without HIV will be enrolled in dose cohorts of 4, monitored carefully for safety, and undergo intensive PK sampling (18 timepoints over 35 days) for plasma API concentrations using a sensitive, validated assay done under GLP. Intracellular API and TFV-diphosphate concentrations from blood mononuclear cells and lymph node mononuclear cells (in a subset) will be compared to plasma concentrations to address the cell-targeting characteristics of the platform technology. The 3-year proposal includes major milestones and a timeline that will be closely monitored to ensure success of this innovative clinical trial. These human data will form a basis to develop more potent highly active LA antiviral combinations with this platform.